Progress 10/13/15 to 09/30/20
Outputs
Target Audience:1. Scientists studying the genetics of muscle development, nutrition, and meat science. 2. Scientists studying molecular pathways of muscle growth and development. 3. Human health and medical community. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?This project provided opportunities for training graduate and undergraduate students with biomedical lab skills and analysis techniques. How have the results been disseminated to communities of interest?We published the first project in the journal of Animals, which has an impact fact2.323, which means it has a great opportunity to be delivered to animal scientists and researchers. We also represented our work in the Experimental Biology meeting, and Animal Science annual meeting. So our work has been seen by researchers all around the world What do you plan to do during the next reporting period to accomplish the goals?We are proposing new studies that use pigs and sheep as models to detect more molecular and signal transduction pathways of animal growth and meat quality that affected by dietary factors, environment, and genetic background.
Impacts
What was accomplished under these goals?
Our first project was to compare the growth performance, meat quality traits, and molecular pathways of muscle and fat growth between commercial cross-bred pigs and Large Black pigs. We have observed the commercial cross-bred pigs have better growth performance, but low meat quality and weaker muscular mitochondrial metabolism. The RNA sequencing and qPCR data proved that there are some key gene expression levelsthat are different between thecommercial cross-bred pigs and Large Black pigs. These genes that recognized affect skeletal muscle growth, intramuscular lipid metabolism, which lead to the differences in the growth performance and meat quality features. The second project was monitoring the growth and meat quality-related gene and molecular pathways in the finishingsheep model when different levels of crude protein were supplied. We found that the high crude protein level promoted growth performance but inhibited the improvement of the meat quality. The different crude protein treatments affected the gene expression related to adipose tissue metabolism. These results are evidence that in the finishing stage, the muscle growth requires a great amount of supplement of crude protein for available amino acids, and the high level of crude protein is utilized as an extra energy source for muscle growth. But the dietary crude protein doesn't promote the meat quality such as the intramuscular marbling formation.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2020
Citation:
S. Ghnaimawi, J.I. Baum, R. Liyanage, and Y. Huang. Concurrent EPA and DHA Supplementation Impairs Brown Adipogenesis of C2C12 Cells. Front. Genet. 11:531. DOI: 10.3389/fgene.2020.00531
- Type:
Journal Articles
Status:
Published
Year Published:
2020
Citation:
4) J.H. Pan, J. Tang, Y.J. Kim, J.H. Lee, E. Shin, J. Zhao, K. Kim, K.A. Hwang, Y. Huang, and J.K. Kim. IDH2 Deficiency Is Critical in Myogenesis and Fatty Acid Metabolism in Mice Skeletal Muscle. 2020. Int. J. Mol. Sci. 2020, 21(16), 5596; https://doi.org/10.3390/ijms21165596
- Type:
Journal Articles
Status:
Published
Year Published:
2021
Citation:
Y. Wang, K. Thakali, P. Morse, S. Shelby, J. Chen, J. Apple, and Y. Huang. Comparison of Growth Performance and Meat Quality Traits of Commercial Cross-Bred Pigs versus the Large Black Pig Breed. 2021. Animals. 2021, 11(1), 200; DOI: 10.3390/ani11010200
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2020
Citation:
S. Ghnaimawi, Y. Huang, J. Baum. Docosahexaenoic Acid (DHA) but not Eicosapentaenoic Acid (EPA) induces the trans-differentiation of C2C12 into white-like adipocytes phenotype. The FASEB Journal, 34: 1-1. https://doi.org/10.1096/fasebj.2020.34.s1.03296
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2020
Citation:
Y. Huang, S. Shelby, Y. Wang, J. Ge, S. Dridi, PSX-37 Late-Breaking Abstracts: Effects of metformin on body temperature regulation and breast muscle growth in male broiler chickens, Journal of Animal Science, Volume 98, Issue Supplement_4, November 2020, Pages 351352, https://doi.org/10.1093/jas/skaa278.617
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2020
Citation:
Y. Huang, S. Ghnaimawi, Y. Wang, S. Zhang, J. Baum, PSIX-33 Docosahexaenoic Acid (DHA) but not Eicosapentaenoic Acid (EPA) induces the trans-differentiation of C2C12 into white-like adipocytes phenotype, Journal of Animal Science, Volume 98, Issue Supplement_4, November 2020, Pages 309310, https://doi.org/10.1093/jas/skaa278.552
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2020
Citation:
Y. Huang, S. Zhang, Y. Wang, S. Ghnaimawi, J. Baum, PSIX-34 Omega-3 fatty acids are more likely to cause C2C12 cell adipose deposition and decomposition compared with palmitic acid, Journal of Animal Science, Volume 98, Issue Supplement_4, November 2020, Pages 308309, https://doi.org/10.1093/jas/skaa278.551
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Progress 10/01/18 to 09/30/19
Outputs
Target Audience:
Nothing Reported
Changes/Problems:Due to the turnover of personnel of the IACUC on campus andthe divided IACUC teams for agriculture research and biomedical research the in vivo study has been delayed unexpectedly. We are reorganizing our animal model, will consider using the lamb model to replace the cattle model to study tall fescue toxicity and animal growth. and using the chicken and pig models to replace the mouse model to study the effect of certain feed additives on thermogenesis. What opportunities for training and professional development has the project provided?This project provided opportunities for training graduate and undergraduate students with biomedical lab skills and analysis techniques. How have the results been disseminated to communities of interest?Our work has been published on the Discovery, The Student Journal of Dale Bumpers College of Agricultural, Food and Life Sciences. So the readers from the University of Arkansas can get the most updated information of our research. We also represented our work in the Experimental Biology meeting, and Animal Science annual meeting. So our work has been seen by the researchers all around the world What do you plan to do during the next reporting period to accomplish the goals?We are closely monitoring and trying to figure the individual function of n-3 fatty acids on muscle growth. And we are pushing our study to in vivo level.
Impacts
What was accomplished under these goals?
Our study suggested that, in the cell culture level, concomitant treatment with EPA and DHA inhibits myotube formation by targeting myogenesis signature genes such as Myf5, MyoD, MyoG, and Pax7, while also promoting the white adipose-like phenotype, as indicated by up-regulation of white adipose-selective genes such as BMP4, C/EBPα, and IL-6. Our data also indicated that combined supplementation of EPA and DHA is correlated with mitochondrial dysfunction, attributed to a reduction in expression of TFAM, COX7a1, and PGC1α genes regulating mitochondrial biosynthesis and function. Lipid droplet accumulation was associated with a reduced oxygen consumption rate. EPA and DHA supplementation were able to further promote lipid storage over induction differentiation media alone, attributed to the higher number of adipocytes formed. Finally, our study demonstrated that white adipocyte-like subsets could arise from mesenchymal precursors expressing Myf5, previously thought to give rise only to skeletal muscle and brown adipocytes. These observations suggested that excessive maternal exposure to EPA and DHA may potentially be involved in obesity and negatively affect muscle.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2019
Citation:
S. Ghnaimawi, S.Shelby, J.I. Baum, and Y. Huang. Effects of Eicosapentaenoic Acid and Docosahexaenoic Acid onC2C12 Cell Adipogenesis and Inhibition of Myotube Formation. 2019. Animal Cells and Systems. 23 (5), 355-364. DOI: 10.1080/19768354.2019.1661282
- Type:
Journal Articles
Status:
Published
Year Published:
2019
Citation:
H. Wu, S. Dridi, Y. Huang, and J.I. Baum. Leucine Decreases Intramyocellular Lipid Deposition in an mTORC1-independent Manner in Palmitate-treated C2C12 Myotubes. 2019. Am J Physiol Endocrinol Metab. 2019 Nov 26. doi:10.1152/ajpendo.00241.2019.
- Type:
Journal Articles
Status:
Published
Year Published:
2019
Citation:
G. Liu, Y. Huang, F. S. Reis, D. Song, and H. Ni. Impact of Nutritional and Environmental Factors on Inflammation, Oxidative Stress, and the Microbiome. BioMed Research International. Volume 2019. DOI: 10.1155/2019/ 5716241
- Type:
Journal Articles
Status:
Published
Year Published:
2019
Citation:
L. Zhao, Y. Huang, and M. Du. Farm Animals are Important Biomedical Models. Animal Frontiers. 2019. Volume 9, Issue 3, July 2019, Pages 2127. DOI: 10.1093/af/vfz015.
- Type:
Journal Articles
Status:
Published
Year Published:
2019
Citation:
B. Alrubaye, M. Abraha, A. Almansour, M. Bansal, H. Wang, Y.M. Kwon, Y. Huang, B. Hargis, and X. Sun. Microbial metabolite deoxycholic acid shapes microbiota against Campylobacter jejuni chicken colonization. PLoS ONE. 2019. 14(7): e0214705. DOI: 10.1371/journal.pone.0214705
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2019
Citation:
S Shelby, K Thakali, Y Wang, J Apple, Y Huang. PSIV-18 Effect of two dietary crude protein levels on finishing performance, meat quality, and genomic changes of lambs. Journal of Animal Science 97 (Supplement_3), 220-221
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2019
Citation:
Y Wang, K Thakali, S Shelby, J Apple, Y Huang. PSII-22 Comparison of the meat quality traits of crossbred pigs and the Large Black pigs. Journal of Animal Science 97 (Supplement_3), 241-242
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2019
Citation:
S Hemza, J Baum, Y Huang. Concurrent Supplementation of Eicosapentaenoic Acid (EPA) and Docosahexaenoic Acid (DHA) Impairs Brown Adipogenesis and Thermogenic Capacity in C2C12 Myoblasts.The FASEB Journal 33 (1_supplement), 721.5.-721.5
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2019
Citation:
S Shelby, S Ghnaimawi, Ji Chen, Y Huang.PSVIII-2 Effects of metformin on white adipose tissue transdifferentiation. Journal of Animal Science 97 (Supplement_3), 304
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2019
Citation:
S Hemza, J Baum, R Liyanage, Y Huang.PSII-23 Concurrent supplementation of n-3 polyunsaturated fatty acids impairs C2C12 from the acquisition of functional brown adipocyte phenotype by negatively affecting the mitochondrial thermogenesis. Journal of Animal Science 97(Supplement_3):242
- Type:
Other
Status:
Published
Year Published:
2019
Citation:
D Dahlem, Y Huang. The Effects of Eicosapentaenoic Acid (EPA) and Docosahexaenoic Acid (DHA) on Brown Adipogenesis in Stem Cell Culture. Discovery, The Student Journal of Dale Bumpers College of Agricultural, Food and Life Sciences. 20_1 27-32
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Progress 10/01/17 to 09/30/18
Outputs
Target Audience:
Nothing Reported
Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?
Nothing Reported
How have the results been disseminated to communities of interest?
Nothing Reported
What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
Our study demonstrated the effects of Eicosapentaenoic acid (EPA) and Docosahexaenoic Acid (DHA) supplement in C2C12 myogenic differentiation. The key gene expression in myogenesis, adipogenesis, mitochondrial, and peroxisomal biosynthesis in C2C12 cells during myogenic differentiation is affected by EPA and DHA. The myotube formation is inhibited while Polyunsaturated fatty acid (PUFA) treatment upregulates adipogenic gene expression. Strengthened peroxisomal biosynthesis and weakened mitochondrial metabolism and biosynthesis imply that the overdosed PUFAs change energy expenditure, and may cause ROS stress that negatively regulates myogenesis. These indicate that the maternal overdosage of EPA and DHA may influence fetal muscle development, increase intramuscular adipose tissue deposition in offspring, and have a long-term effect on the development of metabolic disease such as obesity and diabetes in adult offspring. The sheep study showed that higher dietary crude protein level promotes growth performance for the finishing sheep while lower crude protein level is beneficial for meat quality, especially when evaluating coloration characteristics in the final product.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2018
Citation:
TY Hsueh, JI Baum, and Yan Huang. Effect of Docosahexaenoic Acid and Eicosapentaenoic Acid on Myogenesis and Mitochondrial Biosynthesis during Murine Skeletal Muscle Cell Differentiation. Frontiers of Nutrition. Volume 2018:5(15)
- Type:
Other
Status:
Published
Year Published:
2018
Citation:
Yan Huang, S Hemza, and JI Baum. EPA and DHA Mediated C2C12 Adipogenic Differentiation Involves Up-regulation of Key Adipogenic Markers and Inhibition of Myotubes Formation. 2018. Journal of Animal Science 96 (suppl_3), 244-244
- Type:
Other
Status:
Published
Year Published:
2018
Citation:
X Wang, JA Apple, and Yan Huang. Inactivation of Gene ?-1, 3-Galactosyltransferase in Bovine Aortic Smooth Muscle Cells Using CRISPR-Cas9. 2018. Journal of Animal Science 96 (suppl_2), 105-105
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2018
Citation:
X Wang, JA Apple, and Yan Huang. Inactivation of Gene ?-1, 3-Galactosyltransferase in Bovine Aortic Smooth Muscle Cells Using CRISPR-Cas9. EB2018, abstract.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2018
Citation:
Yan Huang, X Wang, S Shelby, JA Apple, and K Coffey. Effect of Two Dietary Crude Protein Levels on Finishing Performance, Carcass Characteristics, and Meat Quality of Lambs. ASAS Southern Section 2018, abstract.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2018
Citation:
X Wang, JA Apple, and Yan Huang. Inactivation of Gene ?-1, 3-Galactosyltransferase in Bovine Aortic Smooth Muscle Cells Using CRISPR-Cas9. ASAS Midwest Section 2018, abstract.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2018
Citation:
Yan Huang, S Hemza, and JI Baum. EPA and DHA Mediated C2C12 Adipogenic Differentiation Involves Up-regulation of Key Adipogenic Markers and Inhibition of Myotubes Formation. ASAS 2018, abstract.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2018
Citation:
X Wang, JA Apple, and Yan Huang. Inactivation of Gene ?-1, 3-Galactosyltransferase in Bovine Aortic Smooth Muscle Cells Using CRISPR-Cas9. ASAS 2018, abstract.
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Progress 10/01/16 to 09/30/17
Outputs
Target Audience: Scientists studying the genetics of muscle development, nutrition, and meat science. Scientists studying molecular pathways of muscle growth and development. Human health and medical community. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?These projects providenumerous opportunities for graduate students to participate in molecular biology lab work. The graduate students learned analytical techniques include, but not limited to, CRISPR-Cas9 technique, plasmid transfection, sample collection, mRNA extraction, cDNA reverse transcription, and qPCR technique. These lab skills will benefit students for their future lab work. How have the results been disseminated to communities of interest?Part of the cell culture study has been presented in Experimental Biology Meeting 2017, Chicago, IL. Apr 22-26, and in 2017 ASAS-CSAS Annual Meeting, Baltimore, MD. Jul 08-13, 2017. The genomic modified bovine cell culture study has been summarized in an abstract and submitted to 2018 Midwestern Section of ASAS. What do you plan to do during the next reporting period to accomplish the goals?For the C2C12 cell culture study, we will analyze the protein extraction, histochemical staining, mitochondrial respiration of the samples and publish our results in 2018. For the bovine cell culture study, we will select the transfected cells by anti-biotics to build a stablytransfected cell line.
Impacts
What was accomplished under these goals?
Objective 2. Characterize the cellular and molecular basis of myogenesis. Myogenesis and adipogenesis in muscle is affected by n-3 fatty acids supplement for cell culture Preliminary studies were conducted using a murine in vitro myoblasts model (C2C12) to detect the change of mitochondria biosynthesis, function, and pathways of adipogenesis caused by n-3 fatty acid (EPA and DHA) treatment. EPA and DHA were added into culture media to mimic the maternal over-nutrition during gestation. In the myogenesis, data show fatty acids treatment limits the formation of myotubes, as well as the marker genes expression of myogenesis. Genes expression related to adipogenesis are upregulated by fatty acids treatment. Mitochondrial biosynthesis is inhibited by fatty acids, and it regulators' by analyzing mitochondrial respiration, which shows fatty acids treatment decreases the oxygen consumption rate. Peroxisomes biosynthesis genes were upregulated by fatty acids. In white adipogenesis, key genes related to mitochondrial synthesis and metabolism (especially those related to complexes enzymatic activities) were significantly down-regulated by n-3 fatty acids treatment. While during brown adipogenesis, n-3 fatty acids treatment improves ATP synthase, mitochondrial biogenesis, and brown adipogenesis up-stream regulators gene expression. Gene edition technology in transgenic beef production Galactose-α-1,3-galactose (Alpha-Gal) is a mammalian carbohydrate compound that presents in vibrate animals except for humans or Old World monkeys. Alpha-Gal is synthesized by a glycosylation enzyme α-1,3-Galactosyltransferase (GGTA1). Genetically knocking out GGTA1 in pig protects xenotransplantation from hyperacute rejection We have designed target gRNAs for the bovine GGTA1 gene. Next step is using CRISPR-Cas9 to edit GGTA1 gene in bovine primary cell culture. We used the gRNA design tool and selected 5'-GGCCTGACGGTTTTCGCCGT-3' as the target gRNA sequence from the coding DNA sequence of Bos taurus alpha-galactosyltransferase 1 (glycoprotein). The gRNA was constructed in the pSpCas9 BB-2A-GFP (PX458) vector provided by GenScript USA Inc. Vectors were amplified and transfected into BAOSMC by GenePORTER2 transfection reagent when the cells were 80% confluency. Green fluorescent can be viewed after 24 hours transfection. The transfection efficiency can reach about 70% to 80%. Cells were collected in PBS at pH7.4 after 24 hours transfection. Total protein was extracted then the enzyme-linked immunosorbent assay was used to examine the GGTA1 production. By normalized with the total protein concentration, the GGTA1 protein level in the transfected cells was 17.9 ± 7.25% lower (P< 0.05) than in the control cells, showing a significant inhibition of GGTA1 gene expression in the cells by CRISPR-Cas9 gene edition method. Our preliminary data shows that the gRNA sequence that we chose was suitable for the GGTA1 gene knockout in bovine aortic smooth muscle cells. Moreover, the CRISPR-Cas9 system was proved can be applied in the genome editing of bovine cells. Impact: The study in cell culture shows the affected pathways and key gene expressions in muscle cells during myogenesis and adipogenesis affected by n-3 fatty acids treatment The gene edition in beef cattle may provide allergy-free beef for meat allergy patients, and produce alternative organ source for patients need xenotransplantation
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2017
Citation:
TY Hsueh, X Wang, JI Baum, Y Huang. Effect of Fatty Acids on Myogenesis and Mitochondrial Biosynthesis during Murine
Skeletal Muscle Cell Differentiation. The FASEB Journal 31, 877.11-877.11
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2017
Citation:
TY Hsueh, X Wang, Y Huang. Effect of Fatty Acids on Myogenesis and Mitochondrial Biosynthesis during Murine Skeletal
Muscle Cell Differentiation. 2017 ASAS-CSAS Annual Meeting
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Progress 10/13/15 to 09/30/16
Outputs
Target Audience:1. Scientists studying genetics of muscle development, nutrition, and meat science. 2. Scientists studying molecular pathways of muscle growth and development. 3. Human health and medical community. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?This study and future study focus on molecular pathways and genetic study of skeletal muscle biology. The skills of cell and molecular biology are required for the experiments and analysis. There are huge opportunities for training of undergraduate and graduate students, and also for professional development such aspost-doctoral training. How have the results been disseminated to communities of interest?The preliminary study of this projecthas been summarized to an abstract and submitted to Experimental Biology Meeting 2017. What do you plan to do during the next reporting period to accomplish the goals?We will induce adipogenesis in cell culture and detect the genes that have been affected by fatty acids supplement during adipogenesis. After selecting the key genes in myogenesis and adipogenesis that are regulated by fatty acids treatments, we will try to knockout or overexpress some key genes and check their ability of myogenesis and adipogenesis to confirm their connection to fatty acids.
Impacts
What was accomplished under these goals?
We tried to combine the two goals in one study. We treated C2C12 myoblast cells with fatty acids. Signal transduction pathways and mitochondrial function related with skeletal musclegrowth were investigated. Polyunsaturated fatty acids are important nutrients for human health, especially omega-3 fatty acids such as docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), which have been found to play positive roles in the prevention of various diseases. However, previous studies have reported that DHA and EPA can inhibit myoblast proliferation. In this study, C2C12 myoblasts were cultured toconfluencyand then treated with differentiation medium that contained fatty acids (50µM EPA and DHA). After 72 hours of myogenic differentiation, cell mRNA was collected and gene expression was analyzed by real-time PCR. Microscopy was used to examine cell morphology following treatment with fatty acids. In addition, the effect of DHA/EPA on cellular oxygen consumption was measured using a Seahorse XF24 Analyzer. Cells treated with fatty acids had fewer myotubes formed compared to control cells (Fig. 1). The expression of MRF4 (myogenic regulatory factor 4), MyoD (myogenic differentiation), MyoG (myogenin) and Pax7 (paired box 7), genes related to myogenesis, was significantly lower in cells treated with fatty acids (Fig. 2.A). Genes associated with adipogenesis, aP2 (adipocyte protein 2), c/EBPa (CCAAT/enhancer-binding protein alpha), c/EBPb, PPARg (peroxisome proliferator-activated receptor gamma), CPT1 (carnitine palmitoyltransferase I), and FAT/CD36 (fatty acid translocase), had higher expression after treatment with fatty acids (Fig. 2.B). In addition, the mitochondrial biogenesis decreased with lower Mfn2 (mitofusin 2),ERRa(estrogen relatedreceptor alpha), TFAM (mitochondrial transcription factor A), and PGC1a (peroxisome proliferator-activated receptor gamma coactivator 1-alpha) expression and lower mtDNA/nDNA ratio in cells treated with fatty acids compared with control cells (Fig. 2.C and E). However, the expression of PMP70 (70-kDa peroxisomal membrane protein), Pex2 (peroxisomal biogenesis factor 2), and Pex19, genes related to peroxisome biosynthesis, was higher in cells treated with fatty acids (Fig. 2.D). Moreover, fatty acid treatment reduced oxygen consumption rate under oligomycin-inhibited (reflecting proton leak), and uncoupled conditions (Fig. 2.F). Our preliminary data implies that fatty acids might reduce myogenesis and increase adipogenesis in myotube formation. Fatty acids may also decrease cell metabolism by reducing mitochondrial biogenesis as well as respiration rate. These data suggest that high-fat diet or supplement may affect myotube formation during myogenesis in fetal stage and regeneration of skeletal muscle in adults via impaired mitochondrial metabolism.
Publications
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